乳腺导管内乳头状肿瘤187例临床病理及免疫组织化学特征

目的 探讨乳腺导管内乳头状肿瘤(IDPN)的诊断方法和标准.方法 收集187例IDPN患者的临床和病理资料,结合目前认可的2003年WHO乳腺和女性生殖系统肿瘤病理学和遗传学分类标准、Page等和Tavassoli的诊断标准,对其形态学特点进行分析,并对其中53例行CD10、p63、CK14、CK5/6、CK7、乳珠蛋白-1(MGB1)及p53免疫组织化学EnVision法染色分析.结果 187例IDPN患者中导管内乳头状瘤(IDPMa) 128例,不典型导管内乳头状瘤(A-IDPMa) 16例,导管内乳头状癌(IDPCa) 43例.IDPN在形态学上表现为不同程度的上皮细胞和间质增生,以及继发病变等,这些使病灶呈现异常复杂的多样性.免疫组织化学肌上皮标记(CD10和p63)染色在IDPMa、A-IDPMa及IDPCa的表达依次减少,组间比较差异均有统计学意义(均P＜0.001).基底型角蛋白(CK5/6和CK14)染色显示良性病变的表达呈镶嵌状阳性表达,在A-IDPMa的不典型区和IDPCa中表达明显减少或缺如,两者相比差异有统计学意义(P＜0.001).腺腔上皮标志物CK7染色各组间比较差异无统计学意义(P=0.06).MGB1在IDPCa组染色明显减少(P值分别为0.002和0.007),p53染色各组均呈阴性.结论 IDPN是一组组织学改变复杂的疾病,应注意其诊断标准的掌握.肌上皮、基底型角蛋白和腺腔上皮标志物联合应用在该组复杂病变中有很好的诊断和鉴别诊断价值.     

肿瘤增殖和凋亡相关因素在胃癌演进过程中的作用

目的　探讨肿瘤增殖和凋亡相关因素在胃癌发生、发展中的相互作用。方法　采用免疫组化双重染色法对 15 6例正常胃黏膜、不典型增生及胃癌标本分别进行EGFR、Ki 6 7、bcl 2和p5 3的染色。分析两个增殖相关因素 (EGFR、Ki 6 7)之间和凋亡相关因素 (bcl 2、p5 3)之间的相关性。 结果　 (1)上述 4者在正常胃黏膜中均呈阴性表达 ,EGFR在重度不典型增生时呈第一高峰 ,Ⅰ级癌时表达下降 ,Ⅱ～Ⅲ级癌时呈现第二高峰 ,Ki 6 7在胃癌中的表达率 (76 6 % )高于不典型增生 (4 1 4 % ) (P <0 0 1) ,bcl 2在重度不典型增生和Ⅰ级癌中的表达高于轻、中度不典型增生和Ⅱ～Ⅲ级癌 (P <0 0 5 ) ,p5 3在胃癌中的表达高于不典型增生 ,且表达率随分化程度的降低而升高 (P <0 0 5 )。 (2 )EGFR、Ki 6 7和 p5 3的表达与胃不典型增生程度、胃癌分化程度、组织学类型有关 ,而且 ,EGFR和P5 3与浆膜浸润和淋巴结转移有关 ,Ki 6 7虽未达统计学标准 ,但也有此倾向。 (3)EGFR和Ki 6 7表达呈正相关 ,而bcl 2和 p5 3表达呈负相关 (P <0 0 1)。结论　EGFR和Ki 6 7在胃癌发生、发展中有协同作用 ,可作为反映胃癌生物学行为的指标之一。bcl 2和p5 3分别作用于胃癌发生、发展的不同阶段 ,有助于胃癌的早期诊断和预后判断。     

子宫内膜腺癌组织中cyclinD1、PCNA和Ki-67的表达

目的　探讨cyclinD1、PCNA和Ki 6 7在子宫内膜腺癌组织中的表达及其意义。 方法　采用免疫组化S P法检测正常增生期子宫内膜 10例、单纯性增生 30例、复杂性增生 30例、非典型增生 30例和子宫内膜腺癌 4 7例组织中cyclinD1、PCNA和Ki 6 7的表达。结果　cyclinD1在增生期子宫内膜组织中未见阳性表达 ,在单纯性增生、复杂性增生、非典型增生和子宫内膜腺癌组织中的阳性表达率分别为 13 3%、16 7%、30 0 %和 5 7 8%。非典型增生和子宫内膜腺癌组织阳性表达率高于增生期宫内膜、单纯性和复杂性增生 (P <0 0 5 ) ,cyclinD1蛋白表达与子宫内膜腺癌临床分期、肌层浸润和淋巴结转移呈正相关 (P<0 0 5 ) ,而与肿瘤的分化程度无关 (P >0 0 5 )。PCNA和Ki 6 7在非典型增生组标记指数 (LI)高于增生期宫内膜、单纯性和复杂性增生组 ,子宫内膜腺癌组高于非典型增生组 (P <0 0 5 ) ,PCNA和Ki 6 7LI与肿瘤的分化程度、临床分期、肌层浸润和淋巴结转移呈正相关 (P <0 0 5 )。PCNA和Ki 6 7在cyclinD1阳性表达组的LI高于阴性组 (P <0 0 5 )。结论　cyclinD1蛋白过度表达可能在子宫内膜癌的发生、发展中起重要作用 ,其作用途径可能是通过促进细胞增殖而实现的。     

结直肠癌中cyclin D1和p27蛋白的表达及其意义

目的　研究cyclinD1和p2 7蛋白在结直肠癌发生、发展中的作用及其与结直肠癌临床病理特征关系。 方法　收集5 8例手术切除的结直肠癌标本 ,同时取距癌灶 >5cm的正常组织 ,应用免疫组化S P法检测癌组织及正常组织中cyclinD1和p2 7蛋白的表达。结果　cyclinD1蛋白在结直肠癌的表达阳性率为 5 5 17%,正常组织无表达 (P <0 0 1) ;cyclinD1蛋白的表达阳性率在 6 0岁以上年龄组高于 6 0岁以下年龄组 (P <0 0 5 ) ;cyclinD1蛋白的表达与肿瘤组织分化程度负相关 (P <0 0 1)。p2 7蛋白在结直肠癌的表达阳性率为 5 5 17%,在结直肠正常组织的表达阳性率为 96 5 5 %(P <0 0 1) ;p2 7蛋白的表达与肿瘤组织分化程度负相关 (P <0 0 1)。cyclinD1和p2 7蛋白在结直肠癌的表达呈正相关 (r =0 5 82P <0 0 1)。 结论　cyclinD1蛋白过表达与 p2 7蛋白失活可加速细胞周期转化 ,促进结直肠癌的发生 ,cyclinD1和 p2 7蛋白的检测可作为评价结直肠癌恶性程度和判断预后的重要指标。     

新疆维吾尔族与汉族结直肠癌组织P53表达的差异

目的　研究新疆维吾尔族 (维族 )和汉族、浙江汉族结直肠癌组织中 p5 3、Ki 6 7、COX 2蛋白的表达 ,探讨其表达上的差异。方法　应用免疫组化S P法对 14 4例结直肠癌组织 (新疆维族 38例和汉族 5 2例、浙江汉族 5 4例 )中p5 3、Ki 6 7、COX 2蛋白表达进行检测。结果　 3组结直肠癌患者性别构成差异无统计学意义。维族结直肠癌发病年龄早于两组汉族 (浙江汉族、新疆汉族和维族病人的年龄分布分别为 5 7 7± 12 6、5 7 0± 11 5、5 1 4± 11 2岁 ,经方差分析 ,F =3 5 2 0 ,P <0 0 5 )。组织学类型 :浙江汉族病人管状腺癌占 82 7% ,新疆汉族和维族病人乳头状腺癌所占的比例高于浙江汉族 ,3组间差异有显著性(P <0 0 5 )。结直肠癌间分化程度 :新疆汉族高于其他两组 (P <0 0 1)。调整年龄、性别因素后经多因素Logistic回归模型分析 ,民族与结直肠癌p5 3蛋白的阳性表达有关。维族 p5 3阳性率高于汉族 ,比值比为 2 4 92 (95 %CI 1 0 4 6～ 5 939)。新疆汉族和维族结直肠癌与Ki 6 7和COX 2蛋白表达无相关性。结论　维族结直肠癌 p5 3蛋白表达高于汉族病人 ,其意义值得进一步探讨。     

肺癌中P63与P53、E-cadherin、Ki-67表达的比较

目的　比较 p6 3及 p5 3、E cadherin(E cad)、Ki 6 7在肺癌中的表达 ,以了解在不同组织类型肺癌发生发展过程中 ,p6 3与抑癌基因 (p5 3)突变、上皮分化标志基因 (E cad)失活及细胞增殖标志基因 (Ki 6 7)激活有无相关性。方法　采用免疫组化S P法分别检测 6 1例原发性肺癌中 p6 3、p5 3、E cad和Ki 6 7的表达情况。 结果　p6 3在肺鳞癌中阳性率为 10 0 0 % ,而在其他组织类型肺癌中 p6 3基本不表达 ,差异有显著性 (P <0 0 5 ) ;在不同分化程度的肺鳞癌中 p6 3、p5 3的表达差异有显著性 (P<0 0 5 ) ,E cad、Ki 6 7的表达差异无显著性 (P >0 0 5 ) ;E cad的表达在小细胞肺癌与肺鳞癌和肺腺癌之间差异有显著性 (P <0 0 5 ) ;Ki 6 7的表达在各种组织类型肺癌之间差异有显著性 (P <0 0 5 ) ;在不同分化程度鳞癌中 p6 3与E cad的表达呈负相关(P <0 0 5 )。结论　p6 3可作为鳞状上皮源性肿瘤标记物 ,是判断鳞状细胞癌的增殖和分化有意义的指标 ,并可作为鉴别分化差的鳞癌和腺癌、小细胞癌的指标。     

细胞周期调控基因在鼻咽癌发生中的表达

目的　研究 p16、Rb蛋白在各型鼻咽粘膜中的表达及意义。 方法　选择鼻咽粘膜慢性炎、癌前病变及鼻咽癌组织各 30例 ,采用免疫组化LSAB法检测p16、Rb蛋白在上述组织中的阳性表达率。结果　在鼻咽粘膜慢性炎、癌前病变及鼻咽癌组织中 p16蛋白的阳性表达率分别为 10 0 % (0 / 30 )、6 0 % (18/ 30 )及 2 6 .7% (8/ 30 ) ;Rb蛋白的阳性表达率为 76 .7%(2 3/ 30 )、5 6 .7% (17/ 30 )及 33 .3 % (10 / 30 )。经统计学分析 :p16、Rb蛋白在鼻咽粘膜慢性炎症中的阳性表达率高于癌前病变组织 ,差异有显著性 (P <0 .0 1和P <0 .0 5 ) ;p16蛋白在鼻咽癌组织中的表达较癌前病变组织显著下调 (P <0 .0 1) ,而Rb蛋白在两者间的差异无显著性 (P >0 .0 5 )。p16、Rb蛋白的阳性表达率在炎症及癌前病变组织中呈负相关性 (P <0 .0 1) ,而在鼻咽癌组织中二者则无相关性。结论　p16基因的畸变在鼻咽癌的发生中具有直接的促成作用 ;而Rb基因的变异则是鼻咽粘膜癌变的早期事件 ,对鼻咽癌的发生起着间接的促进作用     

宫颈鳞状细胞癌HPV16/18、p53、p21表达及意义

目的　探讨人乳头状瘤病毒 (HPV) 16型、18型及 p5 3、p2 1基因蛋白的表达情况以及与宫颈癌的关系。 方法　应用免疫组化二步法检测 5 0例宫颈鳞癌、4 0例正常宫颈黏膜中HPV16、HPV18、p5 3、p2 1的表达。肿瘤组分 <6 0岁和≥ 6 0岁 2个年龄组 ,观察HPV感染情况。结果　宫颈鳞癌中HPV16、HPV18、p5 3、p2 1表达分别为 4 8%、2 0 %、5 4 %和 5 0 % ,而正常宫颈黏膜中的表达分别为 10 %、0、0、10 % ,两者经统计学比较差异有显著性 (P <0 0 1) ;<6 0岁组和≥ 6 0岁组HPV16阳性率分别为 84 2 %和 2 5 8% ,年青组明显高于年老组 ,经统计学比较差异有显著性 (P <0 0 1)。结论　 (1)宫颈鳞癌的发生与HPV16、HPV18感染有密切关系 ,提示检测宫颈HPV16、HPV18感染情况对于宫颈鳞癌的随访和早期诊断有着重要的参考价值。 (2 )提示宫颈黏膜在HPV感染后 ,可能在p5 3、p2 1多种癌基因的共同作用导致宫颈鳞癌的发生发展。     

乳腺导管内乳头状肿瘤的形态学和免疫表型特征

目的观察乳腺导管内乳头状肿瘤（intraductal papillary neoplasms,IDPN）的形态学和免疫表型特征,并探讨其诊断。方法根据WHO分类（2003）和Tavassoli等（1999、2003）有关标准,筛选出乳腺导管内乳头状肿瘤41例,对上述病例进行HE形态观察和免疫组化检测,选用的一抗有CK5、CK34βE12、CK8、CD34、SMA、p63、CD10。结果（1）导管内乳头状瘤19例,镜检为扩张的导管或小管内含纤维-脉管轴心的乳头状病变,由腺（系）上皮和肌上皮覆盖轴心形成双层构型。其中16例伴腺（系）上皮普通性增生,2例〈10％的乳头覆盖上皮呈不典型增生（AH）改变。腺（系）上皮CK5 17例（89．5％）中～强阳性;CK34βE1216例（84．2％）中～强阳性。肌上皮SMA、p63、CD10均阳性。（2）非典犁导管内乳头状瘤4例,3例伴发于囊内乳头状癌,1例作为主要诊断。表现为≥10％而〈90％的乳头覆盖上皮呈AH改变。其AH区域CK5均阴性,3例CK34βE12阴性～弱阳性。（3）导管内乳头状癌21例,其中①2例仅表现为≥90％的乳头区肌上皮层完全缺乏;②1例仪表现为〉90％的乳头覆盖上皮呈低级导管原位癌（DCIS）样改变;③8例兼具上两种特征;④10例乳头覆盖上皮呈中～高级别DCIS样改变。符合②及③者CK5均阴性,CK34βE12 6例（66．7％）阴性～弱阳性;符合④者CK5、CK34βE12 8例（80％）均阴性～弱阳性。结论IDPN是一组良恶性不同的病变,各自的HE形态和免疫表型均有差异,应采用HE形态和免疫组化相结合的综合标准进行诊断。     

声带早期癌DNA分析及p53、Ki-67和bcl-X的表达

目的　以声带原位癌和早期微小浸润癌为主要研究对象 ,探讨声带从良性到恶性病变各阶段中DNA倍体及基因表达的变化 ,并结合其临床生物学行为 ,深入了解其实质。方法　对 18例声带肿瘤性病变做激光扫描细胞DNA分析并进行随访。对 6 2例声带病变 ,以声带原位癌 (CIS)和早期微小浸润癌 (EMIC)为主 ,与声带浸润性癌、声带息肉分别分为 3组 ,检测p5 3、Ki 6 7和bcl X的基因表达并作各组间对比。 结果　DNA倍体分析表明CIS、EMIC和浸润性癌不同 ,前两者几乎都是二倍体 ,而后者 90 %为异倍体 ;随访结果示CIS、EMIC病人无 1例死于肿瘤 ;还发现DNA二倍体和有DNA凋亡峰的肿瘤患者预后好 ,而肿瘤呈异倍体的患者预后差。免疫表型 :p5 3蛋白在声带肿瘤性病变中表达异常高 ;86 %的CIS、EMIC和 91%的浸润性癌都表达p5 3蛋白 ,阳性指数分别为 2 35和 2 2 6 ,同时此两组的Ki 6 7的阳性平均指数也分别为 2 9%和 2 7% ,与声带息肉差异有显著性 (P <0 0 1)。另一方面 ,bcl X的表达从良性病变到恶性病变呈递减态势 ,以浸润性癌中下降最明显 ,与息肉病变差异有显著性 (P =0 0 0 2 )。结论　声带良性病变、CIS、EMIC及浸润性癌在基因表达和DNA倍体上表现有所不同 ,提示它们各自之间有本质的差别 ,声带原位癌的表现界于良恶     

Co-ordination of cough and swallow in vivo and in silico

Coughing and swallowing are airway-protective behaviours. The pharyngeal phase of swallowing prevents aspiration of oral material (saliva, food and liquid) by epiglottal movement, laryngeal adduction and clearing of the mouth and pharynx. Coughing is an aspiration-response behaviour that removes material from the airway. Co-ordination of these behaviours is vital to protect the airway from further aspiration-promoting events, such as a swallowing during the inspiratory phase of coughing. The operational characteristics, primary strategies and peripheral inputs that co-ordinate coughing and swallowing are unknown. This lack of knowledge impedes understanding and treatment of deficits in airway protection, such as the co-occurrence of dystussia and dysphagia common in Parkinson's and Alzheimer's diseases, as well as stroke.     

ALA和HMME水凝胶栓剂的制备和体内外释药研究

目的 制备光敏剂5-氨基酮戊酸(ALA)和血卟啉单甲醚(HMME)水凝胶栓剂,评价其对直肠肿瘤组织的光敏剂递送效率.方法 将皮下移植人直肠癌细胞SW837的BALB/c小鼠随机分为水凝胶栓剂直肠局部给药组、皮肤局部给药组、瘤内注射给药组和静脉注射给药组.用荧光光谱仪测量直肠壁、皮肤和皮下肿瘤中原卟啉(PpⅨ)和HMME的浓度,荧光光谱系统测定相应的光敏剂分布情况.结果 ALA水凝胶栓剂直肠局部给药组的PpⅨ浓度分别是皮肤局部给药组的9.76倍(1 h)和5.80倍(3 h),差异均具有统计学意义(均P＜0.05).皮肤局部给药后2h,ALA在肿瘤组织内达到最大穿透深度(3～6 mm).而HMME水凝胶栓剂直肠局部给药后,直肠壁中的HMME浓度极低,且皮肤局部给药后的最大肿瘤穿透深度不足2 mm.结论 与皮肤相比,ALA更易穿透黏膜屏障,以水凝胶栓剂形式直肠局部给药有望成为ALA用于光动力疗法治疗直肠癌的一种给药方式.     

Incorporation and Disappearance of Sulfate in Different Regions of Mouse and Rat Costal Cartilage in vivo and in vitro

A rapid and simple method for liquid scintillation counting of ³³S-sulfate in biological tissues is described. Sulfate incorporation per mg dry weight into selected lateral, medial and intermediate regions of ribs was studied using costal cartilage from rats and mice. In vitro, cartilage pieces embracing the osteochondral junction had 2–4 times larger incorporation rate than the remaining homogeneous parts of the ribs, both if entire rib cages or stripped and diced cartilage were incubated. After in vivo injection to rats and mice the incorporation rates of sulfate into the region of the osteochondral junction was 2.6 times that found in “resting” cartilage in rats and 4.4 times in mice. The ³⁵S-sulfate uptake in the ribs per mg dry cartilage diminished in the caudal direction. Growth of the chest cage was mainly a longitudinal growth of the bony segments. Marked differences in the disappearance rates of previously incorporated sulfate were found in different regions along the rib. Rapid sulfate disappearance was found in the bony segment which, however, had incorporated only one seventh of the amount per mg taken up by “resting” cartilage. The “resting” cartilage segments exhibited a steady slow disappearance rate for sulfate and the osteochondral junction region consisting of several differently active tissues showed a rapid sulfate disappearance in the beginning followed by a slow clearance after 2 weeks similar to that of “resting” cartilage. The necessity of careful selection of costal cartilage samples with respect to regional differences is emphasized.     

Characteristics of Tremor in Normal Subjects and in the Diagnosis and Treatment of Parkinsonism

Tensotremorography was used to record voluntary forces and to study the characteristics of the involuntary and voluntary components of isometrically recorded hand strength. The frequency ranges for changes in the spectral density of oscillations recorded here supported the existence of two suprasegmental systems associated with voluntary control and continuous regulation of force maintaining or holding a posture. Cross-correlation analysis of hand force maintained in conditions of visual feedback in normal conditions and in conditions of central disorders of the movement control system is presented.     

Expression of EpCam and villin in Barrett's esophagus and in gastric cardia

In the current study we aimed to clarify the potential of EpCAM and villin as in vivo biomarkers for both Barrett esophagus (BE)-associated neoplasia and BE versus cardiac mucosa. Immunohistochemical staining in BE with various degrees of intraepithelial neoplasia (IN), Barrett carcinoma (BC) and in normal cardiac mucosa (CM) revealed a lack of EpCam and villin in squamous esophageal epithelium. All specimens of IN and BC showed EpCam with varying staining intensities. In 57% of CM samples a weak signal was detected; the remainder displayed strong EpCam expression. Villin was found in 97% of BE specimens and in all those with IN; 37% of BC and 75% of CM specimens were~also positive. We conclude that expression of EpCam and villin differs only between squamous epithelium and BE. Determination of these proteins does not allow discrimination between different degrees of neoplasia or between esophageal intestinal metaplasia and cardiac mucosa.     

Expression of desmin and myoglobin in rhabdomyosarcomas and in developing skeletal muscle

Immunohistochemical staining for desmin and myoglobin was investigated in 35 rhabdomyosarcomas from young people and in skeletal muscle from 16 human fetuses of known gestational age. Twenty-nine of the rhabdomyosarcomas expressed desmin but six undifferentiated or poorly-differentiated tumours were desmin negative. Of the desmin positive cases, most undifferentiated or poorly-differentiated sarcomas expressed desmin alone (12/35). Tumours with increasing rhabdomyoblastomic differentiation co-expressed myoglobin (9/35) and well-differentiated examples also contained cross-striations (7/35). Skeletal muscle from fetuses aged 8 weeks or less consisted mainly of primitive desmin negative round cells. As the cells began to differentiate they quickly expressed desmin and, at approximately 10 weeks, myoglobin was expressed and cross-striations were seen. The combined results strengthen the view that desmin (within a strictly defined context of round cell tumours in young people) is a reliable marker for rhabdomyoblastic differentiation. Support is also given to the notion that very primitive rhabdomyosarcomas may be desmin-negative, although the difficulties of establishing firm diagnoses for some of these tumours is emphasized.     

Fine structure of bovine morulae and blastocysts in vivo and in vitro

 The ultrastructure of bovine morulae and blastocysts developed from in vitro-matured and -fertilized oocytes in a serum-supplemented medium was compared with that of morulae and blastocysts collected non-surgically from superovulated cows. In the in vivo-derived morulae, two characteristic cells types could be identified by the electron-density of their cytoplasm and by their ultrastructural features. One type appeared light in color with low electron-dense cytoplasm. These cells were located in the peripheral layer of the cluster of blastomeres, possessed numerous cellular organelles such as mitochondria and Golgi apparatus and had microvilli projecting into the perivitelline space. The other cell type was distinguished by cytoplasm that stained more densely than that of the lighter-appearing cells. The darker-appearing cells generally possessed fewer organelles than the lighter cells, but many lysosome-like structures were present in the cytoplasm. The in vitro-developed morulae also contained two types of cells similar to those observed in the in vivo morulae. However, most of the in vitro-developed cells possessed numerous lipid droplets and contained fewer lysosome-like structures than the cells of the in vivo-derived morulae. The blastocysts, both in vivo and in vitro, showed a clear differentiation of trophoblast cells and inner cell mass (ICM)-cells. In the in vivo-derived blastocyst, the apical membrane of trophoblast cells was covered with large, numerous microvilli and well-developed junctional complexes were observed. Lipid droplets were present in the cytoplasm of trophoblast and ICM-cells but were not abundant. In vitro-developed blastocysts showed less well-developed junctional complexes between trophoblast cells, less well-developed apical microvilli on the trophoblast cells, and contained large numbers of lipid droplets. This accumulation of lipid droplets was higher in the trophoblast cells than in the ICM-cells. The zonae pellucidae of in vitro-developed embryos were thinner than that of the in vivo-derived embryos. This study demonstrates conspicuous differences in the ultrastructural features between the in vivo-derived and in vitro-developed embryos, suggesting that the ultrastructure may reflect the various physiological anomalies observed in previous studies. Accepted: 29 October 1998